Orally administered Streptococcus thermophilus YIT 2001 is a vehicle for the delivery of glutathione, a reactive reduced thiol, to the intestine.

AIMS: We aimed to analyze the behavior of cellular glutathione of Streptococcus thermophilus strain YIT 2001 (ST-1) in the gastrointestinal environment to understand how orally administered glutathione in ST-1 cells is delivered stably to the intestine in a reactive form, which is essential for its systemic bioavailability against lipid peroxidation. METHODS AND RESULTS: Intracellular glutathione was labeled with L-cysteine-containing stable isotopes. ST-1 cells from fresh culture or lyophilized powder were treated with simulated gastric and intestinal juices for 60 min each. The release of intracellular glutathione in digestive juices was quantified via LC-MS/MS. Most of the cellular glutathione was retained in the gastric environment and released in response to exposure to the gastrointestinal environment. During digestion, the membrane permeability of propidium iodide increased significantly, especially when cells were exposed to cholate, without change in the cell wall state. CONCLUSIONS: ST-1 cells act as vehicles to protect intracellular reactive components, such as glutathione, from digestive stress, and release them in the upper intestine owing to the disruption of membrane integrity induced by bile acid.

Persistence of gut dysbiosis in individuals with anorexia nervosa.

Recent evidence suggests a crucial role of the gut microbiota in the pathogenesis of anorexia nervosa (AN). In this study, we carried out a series of multiple analyses of the gut microbiota of hospitalized individuals with AN over three months using 16S or 23S rRNA-targeted reverse transcription-quantitative polymerase chain reaction (PCR) technology (YIF-SCAN®), which is highly sensitive and enables the precise quantification of viable microorganisms. Despite the weight gain and improvements in psychological features observed during treatment, individuals with AN exhibited persistent gut microbial dysbiosis over the three-month duration. Principal component analysis further underscored the distinct microbial profile of individuals with AN, compared with that of age-matched healthy women at all time points. Regarding the kinetics of bacterial detection, the detection rate of Lactiplantibacillus spp. significantly increased after inpatient treatment. Additionally, the elevation in the Bifidobacterium counts during inpatient treatment was significantly correlated with the subsequent body weight gain after one year. Collectively, these findings suggest that gut dysbiosis in individuals with AN may not be easily restored solely through weight gain, highlighting the potential of therapeutic interventions targeting microbiota via dietary modifications or live biotherapeutics.

Metabolomics profile of Japanese female patients with restricting-type anorexia nervosa.

In this study, the serum metabolic profiles of 10 female patients with restricting type anorexia nervosa (ANR) were compared to those of 10 age-matched healthy female controls. While the levels of amino acids were lower among the patients than among the controls, the levels of uremic toxins, including p-cresyl sulfate (PCS), indole-3-acetic acid, and phenyl sulfate, were higher in ANR patients. The serum PCS levels correlated positively with the abundance of the Clostridium coccoides group or the C. leptum subgroup in the feces of patients, but not in those of controls. Collectively, these results indicate that the serum metabolic profiles of patients with ANR differ from those of healthy women in terms of both decreased amino acid levels and increased uremic toxins. Gut microbes including C. coccoides or C. leptum may be involved in such an increase in uremic toxins.

Safety assessment of the candidate oral probiotic Lactobacillus crispatus YIT 12319: Analysis of antibiotic resistance and virulence-associated genes.

Lactobacillus crispatus YIT 12319 (LcY) was isolated from the oral cavity of a healthy subject as a new candidate probiotic with potential benefits for oral health. As a safety assessment of LcY, we performed an antibiotic susceptibility test and virulence-associated gene analysis using a draft genome sequence. Susceptibility to 15 antibiotics was analyzed according to the standard method of the International Dairy Federation/International Organization for Standardization, as recommended by the European Food Safety Authority. The results showed that the minimum inhibitory concentrations of LcY were not higher than those of other L. crispatus strains, which have not acquired resistance to any antibiotics, suggesting that LcY had no externally acquired transmissible antibiotic resistance genes. Analysis of virulence-associated genes using the draft genome of LcY found that there were fewer potential virulence-associated genes in LcY than in other probiotics. These findings suggest that LcY could be a candidate probiotic based on its safety profile.

Intake of ß-cryptoxanthin with fat-containing food increases ß-cryptoxanthin serum level and palmar yellowness in healthy adults.

Epidemiological and clinical studies have suggested that ß-cryptoxanthin (ß-CX) has health benefits in humans. To understand the health benefits of ß-CX, it is important to examine its in vivo kinetics and identify a convenient noninvasive biomarker for serum ß-CX level. However, to date, there have been few studies of ß-CX kinetics in humans. We hypothesized that simultaneous consumption of fat-containing food would stimulate absorption of ß-CX. We conducted 2 in vivo kinetics studies, one after a single intake and the other after daily intake of ß-CX in healthy adults, to examine whether simultaneous consumption of fat-containing food stimulates absorption of ß-CX and whether palmar b* value (yellowness) is a suitable biomarker. After a single intake of 1.1 or 2.2 mg of ß-CX, the serum level increased dose-dependently and returned to the baseline level after 14 to 17 days. The simultaneous consumption of fat-containing food enhanced the absorption of ß-CX by 1.8-fold. During daily intake of 2.0 mg/day ß-CX with fat-containing food for 12 weeks, both serum ß-CX level and palmar b* value measured with a colorimeter increased continuously. After intake was halted, both serum ß-CX level and palmar b* value decreased. There was a positive correlation between serum ß-CX level and palmar b* value during the trial (R = 0.55, P < .001). These results suggest that intake of ß-CX with fat-containing food stimulates the absorption of ß-CX and increases palmar yellowness.

Independent and Interactive Effects of Habitually Ingesting Fermented Milk Products Containing Lactobacillus casei Strain Shirota and of Engaging in Moderate Habitual Daily Physical Activity on the Intestinal Health of Older People.

Infrequent bowel movements decrease the number of beneficial bacteria in the human intestines, thereby potentially increasing the individual's risk of colorectal cancer. The correction of such bowel problems could therefore make an important contribution to improving population health and quality-adjusted lifespan. We examined independent and interactive effects upon the fecal microbiota of two potentially favorable determinants of intestinal motility: the intake frequency of a fermented milk product containing Lactobacillus casei strain Shirota (LcS) and the quantity/quality of habitual physical activity in 338 community-living Japanese aged 65-92 years. Subjects were arbitrarily grouped on the basis of questionnaire estimates of LcS intake (0-2, 3-5, and 6-7 days/week) and pedometer/accelerometer-determined patterns of physical activity [<7000 and ≥7000 steps/day, or <15 and ≥15 min/day of activity at an intensity >3 metabolic equivalents (METs)]. After adjustment for potential confounders, the respective numbers of various beneficial fecal bacteria tended to be larger in more frequent consumers of LcS-containing products, this trend being statistically significant (mostly P < 0.001) for total Lactobacillus, the Lactobacillus casei subgroup, and the Atopobium cluster; in contrast, there were no statistically significant differences in fecal bacterial counts between the physical activity groups. A multivariate-adjusted logistic regression analysis estimated that the risk of infrequent bowel movements (arbitrarily defined as defecating ≤3 days/week) was significantly lower (P < 0.05) in subjects who ingested LcS-containing products 6-7 rather than 0-2 days/week [odds ratio (95% confidence interval) 0.382 (0.149-0.974)] and was also lower in those who took ≥7000 rather than <7000 steps/day [0.441 (0.201-0.971)] or spent ≥15 rather than <15 min/day of physical activity at an intensity >3 METs [0.412 (0.183-0.929)]. The risk of infrequent bowel movements in subjects who combined 6-7 days/week of LcS with ≥7000 steps/day or ≥15 min/day of activity at >3 METs was only a tenth of that for individuals who combined 0-2 days/week of LcS with <7000 steps/day or <15 min/day at >3 METs. These results suggest that elderly individuals can usefully ingest LcS-containing supplements regularly (≥6 days/week) and also engage in moderate habitual physical activity (≥7000 steps/day and/or ≥15 min/day at >3 METs) in order to enhance their gastrointestinal health.

Effect of Lactobacillus casei strain Shirota-fermented milk on metabolic abnormalities in obese prediabetic Japanese men: a randomised, double-blind, placebo-controlled trial.

An obesity-related prediabetic state is characterised by metabolic abnormalities such as post-glucose load hyperglycaemia and dyslipidaemia and consequently increases the risk for type 2 diabetes and cardiovascular disease. This study aimed to investigate the effects of Lactobacillus casei strain Shirota (LcS) on metabolic abnormalities in obese prediabetic subjects in a randomised, double-blind, placebo-controlled trial. Herein, 100 obese subjects (body mass index ≥25), who had moderate post-load hyperglycaemia (1-hr post-load plasma glucose (PG) levels ≥180 mg/dl during the oral glucose tolerance test), consumed LcS-fermented milk or placebo milk daily for 8 weeks. The post-load PG and fasting blood markers were evaluated. Although post-load PG levels were not significantly different between the groups, 1-hr post-load PG, glycoalbumin, and HbA1c levels decreased at 8 weeks compared with the baseline levels only in the LcS group (p=0.036, p=0.002, and p=0.006, respectively). The reduction in glycoalbumin levels was statistically significantly greater in the LcS group than in the placebo group (p=0.030). Stratified analyses revealed significantly improved 1-hr post-load PG and glycoalbumin levels in the LcS group compared with the placebo group among subjects with severe glucose intolerance (2-hr post-load PG levels higher than the median at baseline; p=0.036 and p=0.034, respectively). In terms of lipidic outcomes, total, low-density lipoprotein, and non-high-density lipoprotein cholesterol levels were significantly lower in the LcS group than in the placebo group (p=0.023, p=0.022, and p=0.008, respectively). These findings suggest that LcS may favourably affect metabolic abnormalities in obese prediabetic subjects, though the effects on glycaemic control may be limited.

Beneficial Effects of Lemon Balm Leaf Extract on In Vitro Glycation of Proteins, Arterial Stiffness, and Skin Elasticity in Healthy Adults.

Glycation, a non-enzymatic glycosylation of proteins, induces tissue damage in association with various diseases and aging phenomena. Pentosidine, an advanced glycation end product, is involved in aging phenomena such as tissue stiffness. In this study, we aimed to find a potent anti-glycation food material and to verify its health benefits by clinical trial. From among 681 hot water plant extracts, lemon balm (Melissa officinalis; LB) leaf extract was selected and revealed to have more potent inhibitory activity for pentosidine formation than a representative anti-glycation agent, aminoguanidine. Rosmarinic acid (RA), a typical polyphenol in Lamiaceae plants, was identified as a major active component in LB extract (LBE). Furthermore, LBE or RA dose-dependently suppressed glycation-associated reactions such as increased fluorescence, yellowing of collagen fiber sheets, and degeneration of the fibrous structure of elastin fiber sheets. An open-label, parallel-group comparative trial was conducted in 28 healthy Japanese subjects aged 31-65 y who consumed LB tea (LB group) or barley tea (Control group) for 6 wk. The LB group showed significant reductions in brachial-ankle pulse wave velocity, reflecting arterial stiffness, and b* (yellow) color values in forearm skin compared with the Control group. A gender-stratified analysis revealed that cheek skin elasticity was significantly improved in the LB group compared with the Control group only in female subjects. It is concluded that the hot water extract of LB leaf has the potential to provide health benefits with regard to glycation-associated tissue damage in blood vessels and skin of healthy adults.

Daily intake of fermented milk with Lactobacillus casei strain Shirota reduces the incidence and duration of upper respiratory tract infections in healthy middle-aged office workers.

PURPOSE: Although several studies have demonstrated the efficacy of probiotics for preventing upper respiratory tract infections (URTIs) in at-risk populations, including children and the elderly, few studies have investigated the efficacy of probiotics in healthy adults living normal, everyday lives. Thus, we tried to evaluate the effects of Lactobacillus casei strain Shirota-fermented milk (LcS-FM) on the incidence of URTIs in healthy middle-aged office workers. METHODS: In a randomized controlled trial, 96 eligible male workers aged 30-49 years consumed LcS-FM containing 1.0 × 1011 viable LcS cells or control milk (CM) once daily for 12 weeks during the winter season. URTI episodes were evaluated by a physician via a questionnaire of URTI symptoms. RESULTS: The incidence of URTIs during the intervention period was significantly lower in the LcS-FM group than in the CM group (22.4 vs. 53.2 %, P = 0.002). The time-to-event analysis showed that the LcS-FM group had a significantly higher URTI-free rate than the CM group over the test period (log-rank test: χ 2 11.25, P = 0.0008). The cumulative number of URTI episodes and cumulative days with URTI symptoms per person was lower in the LcS-FM group, and the duration per episode was shorter. Inhibition of both reductions in NK cell activity in peripheral blood mononuclear cells and increases in salivary cortisol levels was observed in the LcS-FM group. CONCLUSION: The results suggest that the daily intake of fermented milk with LcS may reduce the risk of URTIs in healthy middle-aged office workers, probably through modulation of the immune system.

Prebiotic potential of L-sorbose and xylitol in promoting the growth and metabolic activity of specific butyrate-producing bacteria in human fecal culture.

Dietary low-digestible carbohydrates (LDCs) affect gut microbial metabolism, including the production of short-chain fatty acids. The ability of various LDCs to promote butyrate production was evaluated in in vitro human fecal cultures. Fecal suspensions from five healthy males were anaerobically incubated with various LDCs. L-Sorbose and xylitol markedly promoted butyrate formation in cultures. Bacterial 16S rRNA gene-based denaturing gradient gel electrophoresis analyses of these fecal cultures revealed a marked increase in the abundance of bacteria closely related to the species Anaerostipes hadrus or A. caccae or both, during enhanced butyrate formation from L-sorbose or xylitol. By using an agar plate culture, two strains of A. hadrus that produced butyrate from each substrate were isolated from the feces of two donors. Furthermore, of 12 species of representative colonic butyrate producers, only A. hadrus and A. caccae demonstrated augmented butyrate production from L-sorbose or xylitol. These findings suggest that L-sorbose and xylitol cause prebiotic stimulation of the growth and metabolic activity of Anaerostipes spp. in the human colon.

Effect of probiotic and prebiotic fermented milk on skin and intestinal conditions in healthy young female students.

Although persistent constipation is considered to be associated with skin problems, the supporting evidence is limited. Hence, this study investigated the effects of probiotic and prebiotic fermented milk on skin and intestinal conditions in an open-label trial. Among the 101 Japanese healthy young female students that participated, the 81 subjects in the intake group consumed a bottle of probiotic and prebiotic fermented milk containing Bifidobacterium breve strain Yakult and galactooligosaccharides daily for 4 weeks after a pre-intake period of 4 weeks, while the 20 subjects in the non-intake group did not consume the test beverage. Defecation patterns, skin conditions including hydration levels in the stratum corneum, and urinary phenol and p-cresol levels were evaluated before (baseline) and after intake. All subjects completed the study. No differences in dietary intake and body mass index were observed between both groups during the trial. In the intake group, hydration levels of the stratum corneum, defecation frequency, and feces quantity significantly increased, and urinary phenol and p-cresol levels significantly decreased after intake compared with the corresponding baseline values. However, they did not significantly change in the non-intake group. A significant difference was observed between the intake group and non-intake group in regard to clearness of the skin as assessed by visual analogue scale. Therefore, consecutive intake of probiotic and prebiotic fermented milk might have beneficial effects on the skin that prevent dryness and beneficial effects on the intestinal conditions that stimulate defecation and decrease phenol production by gut bacteria in healthy young women.

Oral Administration of Fermented Soymilk Products Protects the Skin of Hairless Mice against Ultraviolet Damage.

The protective effect of isoflavones on skin damage from ultraviolet (UV) radiation and their bioavailability were investigated in ovariectomized hairless mice fed diets composed of fermented soymilk containing aglycone forms of isoflavones or control soymilk containing glucose-conjugated forms of isoflavones. The erythema intensity of dorsal skin was significantly higher in ovariectomized mice than in sham-operated mice (p < 0.05). The erythema intensity and epidermal thickness of dorsal skin were significantly lower in the fermented soymilk diet group than in the control diet group (each p < 0.05). Levels of cyclobutane pyrimidine dimers in dorsal skin were significantly lower in the fermented soymilk diet group than in the control group (p < 0.05). Serum and dorsal skin isoflavone concentrations were significantly higher in the fermented soymilk diet group than in the soymilk diet group (p < 0.05). These results indicate that oral administration of a fermented soymilk diet increases isoflavone concentrations in the blood and skin, effectively scavenging the reactive oxygen species generated by UV irradiation and exerting an estrogen-like activity, with a consequent protective effect on skin photodamage in hairless mice.

Fermented Milk Containing Lactobacillus casei Strain Shirota Preserves the Diversity of the Gut Microbiota and Relieves Abdominal Dysfunction in Healthy Medical Students Exposed to Academic Stress.

UNLABELLED: Stress-induced abdominal dysfunction is an attractive target for probiotics. To investigate the effects of the probiotic Lactobacillus casei strain Shirota on abdominal dysfunction, a double-blind, placebo-controlled trial was conducted with healthy medical students undertaking an authorized nationwide examination for academic advancement. For 8 weeks, until the day before the examination, 23 and 24 subjects consumed an L. casei strain Shirota-fermented milk and a placebo milk daily, respectively. In addition to assessments of abdominal symptoms, psychophysical state, and salivary stress markers, gene expression changes in peripheral blood leukocytes and composition of the gut microbiota were analyzed using DNA microarray analysis and 16S rRNA gene amplicon sequence analysis, respectively, before and after the intervention. Stress-induced increases in a visual analog scale measuring feelings of stress, the total score of abdominal dysfunction, and the number of genes with changes in expression of more than 2-fold in leukocytes were significantly suppressed in the L. casei strain Shirota group compared with those in the placebo group. A significant increase in salivary cortisol levels before the examination was observed only in the placebo group. The administration of L. casei strain Shirota, but not placebo, significantly reduced gastrointestinal symptoms. Moreover, 16S rRNA gene amplicon sequencing demonstrated that the L. casei strain Shirota group had significantly higher numbers of species, a marker of the alpha-diversity index, in their gut microbiota and a significantly lower percentage of Bacteroidaceae than the placebo group. Our findings indicate that the daily consumption of probiotics, such as L. casei strain Shirota, preserves the diversity of the gut microbiota and may relieve stress-associated responses of abdominal dysfunction in healthy subjects exposed to stressful situations. IMPORTANCE: A novel clinical trial was conducted with healthy medical students under examination stress conditions. It was demonstrated that the daily consumption of lactic acid bacteria provided health benefits to prevent the onset of stress-associated abdominal symptoms and a good change of gut microbiota in healthy medical students.

Intragastric injection of Lactobacillus casei strain Shirota suppressed spleen sympathetic activation by central corticotrophin-releasing factor or peripheral 2-deoxy-d-glucose in anesthetized rats.

Intragastric (IG) administration of probiotic strain Lactobacillus casei Shirota (LcS) decreases the sympathetic nerve outflow of anesthetized rats in a tissue-specific manner. In the present study, we examined the effects of IG administration of LcS on sympathetic activation induced by an intracerebroventricular (ICV) injection of corticotrophin-releasing factor (CRF) and an intravenous (IV) injection of 2-deoxy-d-glucose (2DG) or interleukin (IL)-1ß in urethane-anesthetized rats. The IG administration of LcS differently affected the stimulatory responses of sympathetic nerve outflow to CRF. LcS suppressed the increase in splenic sympathetic nerve activity (Spleen-SNA), induced by central CRF, in a dose-dependent manner; however, it did not alter adrenal sympathetic nervous activity (ASNA). In contrast, LcS did not affect spleen-SNA and ASNA following an IV injection of IL-1ß. On the other hand, IG administration of LcS suppressed the activation of ASNA following an IV injection of 2DG. These findings suggest that the suppression of central CRF-induced sympathetic activation by LcS is tissue-specific. Moreover, it can suppress the 2DG-induced sympathetic activation. Furthermore, we found that stomach-specific vagotomy attenuates the suppressive effect of LcS on CRF-mediated spleen-SNA activation. Thus, the present study suggests that LcS administered to the stomach may act on the afferent vagal nerve and send afferent signals to the brain to regulate efferent SNA induced by sympathetic stimulators.

Intake of a fermented soymilk beverage containing moderate levels of isoflavone aglycones enhances bioavailability of isoflavones in healthy premenopausal Japanese women: a double-blind, placebo-controlled, single-dose, crossover trial.

This study aimed to investigate the bioavailability of serum isoflavones after the intake of soymilk fermented by Lactobacillus casei strain Shirota containing 32.5% isoflavone aglycones (FSM) or placebo soymilk containing no isoflavone aglycones (SM). In a double-blind, placebo-controlled, single-dose, crossover trial, 7 healthy premenopausal Japanese women (mean age: 35.3 ± 11.0) consumed FSM or SM on day 1 and crossed over to the other soymilk after a 6-day washout period. Serum isoflavones in blood samples collected at 0, 1, 2, 3, 4, and 5 hr after intake were analyzed by liquid chromatography coupled with tandem mass spectrometry. The area under the curve (AUC) values for the serum concentrations of genistein and total isoflavones were significantly higher, by about 1.4-fold, up to 5 hr after FSM intake compared with SM intake (each p<0.05), and that of daidzein tended to be higher after FSM intake. In addition, AUC analysis of total isoflavones for individual subjects revealed that 5 out of 7 subjects had higher AUC values after FSM intake compared with SM intake and that the 2 remaining subjects had similar AUC values. These 2 subjects had higher AUC values after SM intake (mean, 2,502 ± 348) than those of the other subjects (mean, 1,158 ± 269). These results indicate that the bioavailability of isoflavones, especially genistein, is enhanced after the intake of FSM containing 32.5% isoflavone aglycones compared with intake of SM containing no isoflavone aglycones and that the enhancement is observed in healthy premenopausal Japanese women whose isoflavone absorption capacity is low after SM intake.

Beneficial effects of citrus juice fermented with Lactobacillus plantarum YIT 0132 on atopic dermatitis: results of daily intake by adult patients in two open trials.

This study aimed to examine whether daily intake of citrus juice containing heat-killed Lactobacillus plantarum YIT 0132 (LP0132-fermented juice) alleviates symptoms of atopic dermatitis. This was a natural extension of our previous study in which LP0132 was shown to enhance IL-10 production in vitro and LP0132-fermented juice was found to alleviate symptoms and enhance quality of life (QOL) in patients with Japanese cedar pollinosis. In two open trials, Trial 1 and Trial 2, 32 and 18 adult patients with mild to moderate atopic dermatitis consumed LP0132-fermented juice for 8 weeks. Skin conditions and QOL were subjectively evaluated using Skindex-16 before intake of the juice (Pre-treatment), 8 weeks after starting intake (Treatment) and 8 weeks after termination of intake (Post-treatment). Blood parameters were also analyzed. Comparison of the Treatment and Post-treatment time points with the Pre-treatment time point revealed significant reductions in the Skindex-16 overall score and the 3 domain subscores (symptoms, emotions, and functioning domains) in both trials. Moreover, blood levels of eosinophil cationic protein (ECP), total immunoglobulin E (IgE) and specific IgEs for Japanese cedar and cypress pollen were significantly attenuated in Trial 2. The findings suggest that daily intake of citrus fermented juice containing heat-killed LP0132 has beneficial effects on symptoms and QOL in patients with mild to moderate atopic dermatitis due to an immunomodulatory effect via attenuation of IgE and ECP.

Gut Dysbiosis in Patients with Anorexia Nervosa.

Anorexia nervosa (AN) is a psychological illness with devastating physical consequences; however, its pathophysiological mechanism remains unclear. Because numerous reports have indicated the importance of gut microbiota in the regulation of weight gain, it is reasonable to speculate that AN patients might have a microbial imbalance, i.e. dysbiosis, in their gut. In this study, we compared the fecal microbiota of female patients with AN (n = 25), including restrictive (ANR, n = 14) and binge-eating (ANBP, n = 11) subtypes, with those of age-matched healthy female controls (n = 21) using the Yakult Intestinal Flora-SCAN based on 16S or 23S rRNA-targeted RT-quantitative PCR technology. AN patients had significantly lower amounts of total bacteria and obligate anaerobes including those from the Clostridium coccoides group, Clostridium leptum subgroup, and Bacteroides fragilis group than the age-matched healthy women. Lower numbers of Streptococcus were also found in the AN group than in the control group. In the analysis based on AN subtypes, the counts of the Bacteroides fragilis group in the ANR and ANBP groups and the counts of the Clostridium coccoides group in the ANR group were significantly lower than those in the control group. The detection rate of the Lactobacillus plantarum subgroup was significantly lower in the AN group than in the control group. The AN group had significantly lower acetic and propionic acid concentrations in the feces than the control group. Moreover, the subtype analysis showed that the fecal concentrations of acetic acid were lower in the ANR group than in the control group. Principal component analysis confirmed a clear difference in the bacterial components between the AN patients and healthy women. Collectively, these results clearly indicate the existence of dysbiosis in the gut of AN patients.

Gel phantom study of a cryosurgical probe with a thermosiphon effect and liquid nitrogen-cooled aluminum thermal storage blocks.

Cryosurgery is a minimally invasive treatment for certain types of cancers. Argon-based cryosurgical devices are available at present, however a large compressed gas cylinder with the pressure of 300 atmospheres is needed. To overcome these drawbacks, we developed a new cryosurgical probe measuring about 50 cm in length with separate lumens inside for liquid and gaseous ethylene to be used as a thermosiphon and liquid nitrogen-cooled aluminum thermal storage blocks. The probe needle was 8 cm in length and 3 mm in outer diameter. To investigate the freezing capabilities of our new cryosurgical system we inserted the needle 5cm into a poly-acrylamide gel phantom warmed to 36.5 ℃. Thermal storage blocks made of aluminum, cooled at -196 ℃ in liquid nitrogen, were attached to the condenser of the probe and replaced with thermal storage blocks every 4 to 5 minutes to compensate for warming. We took digital camera images of the ice ball at the needle and measured the temperature in certain locations of the cryoprobe. Ice ball formation started at one minute after cooling. The sizes (longest diameter × minimum diameter) at 10, 20 and 30 minutes after the start of the procedure were 4.5×2.1, 4.5×3.1 and 4.6×3.7 cm, respectively. During the procedure the minimum temperature of the condenser was -85 ℃ and the needle was -65 ℃. This newly developed compact cryosurgical probe with thermosiphon effect and cooled thermal storage blocks created an ice ball that can be used for cryosurgery within 20 minutes.

Screening of Probiotic Candidates in Human Oral Bacteria for the Prevention of Dental Disease.

The oral cavity in healthy subjects has a well-balanced microbiota that consists of more than 700 species. However, a disturbance of this balance, with an increase of harmful microbes and a decrease of beneficial microbes, causes oral disorders such as periodontal disease or dental caries. Nowadays, probiotics are expected to confer oral health benefits by modulating the oral microbiota. This study screened new probiotic candidates with potential oral health benefits and no harmful effects on the oral cavity. We screened 14 lactobacillus strains and 36 streptococcus strains out of 896 oral isolates derived from healthy subjects. These bacteria did not produce volatile sulfur compounds or water-insoluble glucan, had higher antibacterial activity against periodontal bacteria, and had higher adherence activity to oral epithelial cells or salivary-coated hydroxyapatite in vitro. We then evaluated the risk of primary cariogenicity and infective endocarditis of the selected oral isolates. As a result, Lactobacillus crispatus YIT 12319, Lactobacillus fermentum YIT 12320, Lactobacillus gasseri YIT 12321, and Streptococcus mitis YIT 12322 were selected because they showed no cariogenic potential in an artificial mouth system and a lower risk of experimental infective endocarditis in a rat model. These candidates are expected as new probiotics with potential oral health benefits and no adverse effects on general health.